High-throughput assays probing protein-RNA interactions of eukaryotic translation initiation factors.
Protein-RNA interactions are involved in all facets of RNA biology. The identification of small molecules that selectively block such bimolecular interactions could provide insight into previously unexplored steps of gene regulation. Such is the case for regulation of eukaryotic protein synthesis where interactions between messenger RNA (mRNA) and several eukaryotic initiation factors govern the recruitment of 40S ribosomes (and associated factors) to mRNA templates during the initiation phase. We have designed simple fluorescence polarization-based high-throughput screening assays that query the binding of several translation factors to RNA and found that the mixed inhibitor p-chloromercuribenzoate interferes with poly(A) binding protein-RNA interaction.
Galicia-Vázquez
Gabriela
G
Department of Biochemistry, McGill University, Montreal, Que., Canada H3G1Y6.
Lindqvist
Lisa
L
Wang
Xiaofeng
X
Harvey
Isabelle
I
Liu
Jing
J
Pelletier
Jerry
J
D039642:Eukaryotic Initiation Factors / Q000032:analysis / Q000378:metabolism*; D005454:Fluorescence Polarization / Q000379:methods*; D006801:Humans; D039102:Poly(A)-Binding Proteins / Q000378:metabolism; D012313:RNA / Q000032:analysis / Q000378:metabolism*; D020245:p-Chloromercuribenzoic Acid / Q000737:chemistry
D039642:Eukaryotic Initiation Factors; D039102:Poly(A)-Binding Proteins; D020245:p-Chloromercuribenzoic Acid; D012313:RNA